Binding of a Highly De-N-acetylated Chitosan to Japanese Pheasan Lysozyme as Measured by ^1H-NMR Spectroscopy(Biochemistry & Molecular Biology)

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概要

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• Binding of a highly de-N-acetylated chitosan to Japanese pheasant lysozyme (JPL), which differs from hen egg white lysozyme (HEWL) by nine amino acid substitutions (including Arg114→His), was investigated by ^1H-NMR spectroscopy. The profile of the one-dimensional spectrum of JPL is essentially identical to that of HEWL. Using two-dimensional spectra of JPL and HEWL, several aromatic and aliphatic proton resonances of JPL were assigned by comparison. When a highly de-N-acetylated chitosan (number-average degree of polymerization, about 18 ; degree of acetylation, 0.04), where the N-acetylated units are predominantly surrounded by de-N-acetylated units (a monoacetylated chitosan), was added to the JPL solution, the NMR signals were clearly affected in Trp28 C5H and Ile98 γCH, as in the case of binding to HEWL. The dissociation constant of the monoacetylated chitosan evaluated from the NMR signal responses was calculated to be 0.23 ± 0.05 mм (-31.5kJ/mol), which is similar to that of HEWL (0.11 ± 0.02 mм, -33.3 kJ/mol). Thus, the Arg→His substitution of the 114th amino acid, which participates in sugar residue binding at the right-sided subsite F, did not significantly affect the chitosan binding. In addition, the C2H signal of His114 of JPL was not affected by the chitosan binding. These results suggest that the monoacetylated chitosan binds to subsites E and F through the left-sided binding mode.

• 社団法人日本農芸化学会の論文
• 2001-08-23

著者

• ARAKI Tomohiro

  Department of Bioscience, School of Agriculture, Tokai University

• FUKAMIZO Tamo

  Laboratory of Enzyme System Science, Faculty of Agriculture, Kinki University

• VÅRUM Kjell

  Department of Biotechnology, Norwegian University of Science and Technology

• Fukamizo T

  新潟大学 農学部応用生物化学科

• Fukui T

  Faculty Of Agriculture Kinki University

• Fukamizo Tamo

  Laboratory Of Biophysical Chemistry Faculty Of Agriculture Kinki University

• Kristiansen Are

  Norwegian Biopolymer Laboratory (nobipol) Department Of Biotechnology The Norwegian University Of Sc

• Araki T

  Department Of Bioscience School Of Agriculture Kyushu Tokai University

• Araki Tomohiro

  Department Of Bioscience School Of Agriculture Kyushu Tokai University

• Torikata T

  Department Of Bioscience School Of Agriculture Tokai University

• Torikata Takao

  Laboratory Of Sericultural Chemistry Faculty Of Agriculture Kyushu University

• YAMAGUCHI Tsugihisa

  Laboratory of Enzyme System Science, Department of Food and Nutrition, Kinki University

• ARAKI Tomohiro

  Laboratory of Biochemistry, Faculty of Agriculture, Kyushu Tokai University

• VARUM Kjell

  Norwegian Biopolymer Laboratory (NOBIPOL), Department of Biotechnology, The Norwegian University of

• Varum Kjell

  Norwegian Biopolymer Laboratory (nobipol) Department Of Biotechnology The Norwegian University Of Sc

• Yamaguchi Tsugihisa

  Laboratory Of Enzyme System Science Department Of Food And Nutrition Kinki University

• Varum Kjell

  Norwegian Biopolymer Laboratory (nobipol) Department Of Biotechnology Norwegian University Of Scienc

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